Please use this identifier to cite or link to this item: http://repositorio.unicamp.br/jspui/handle/REPOSIP/199743
Type: Artigo de periódico
Title: Use Of In Vivo And In Vitro Systems To Select Leishmania Amazonensis Expressing Green Fluorescent Protein.
Author: Costa, Solange dos Santos
de Assis Golim, Marjorie
Rossi-Bergmann, Bartira
Costa, Fabio Trindade Maranhão
Giorgio, Selma
Abstract: Various Leishmania species were engineered with green fluorescent protein (GFP) using episomal vectors that encoded an antibiotic resistance gene, such as aminoglycoside geneticin sulphate (G418). Most reports of GFP-Leishmania have used the flagellated extracellular promastigote, the stage of parasite detected in the midgut of the sandfly vector; fewer studies have been performed with amastigotes, the stage of parasite detected in mammals. In this study, comparisons were made regarding the efficiency for in vitro G418 selection of GFP-Leishmania amazonensis promastigotes and amastigotes and the use of in vivo G418 selection. The GFP-promastigotes retained episomal plasmid for a prolonged period and G418 treatment was necessary and efficient for in vitro selection. In contrast, GFP-amastigotes showed low retention of the episomal plasmid in the absence of G418 selection and low sensitivity to antibiotics in vitro. The use of protocols for G418 selection using infected BALB/c mice also indicated low sensitivity to antibiotics against amastigotes in cutaneous lesions.
Subject: Amebicides
Animals
Flow Cytometry
Gentamicins
Green Fluorescent Proteins
Host-parasite Interactions
Leishmania Mexicana
Leishmaniasis, Cutaneous
Luminescent Agents
Macrophages, Peritoneal
Mice
Mice, Inbred Balb C
Organisms, Genetically Modified
Spectrometry, Fluorescence
Leishmania Amazonensis
Geneticin
Green Fluorescent Protein
Macrophage
Rights: aberto
Identifier DOI: 10.3347/kjp.2011.49.4.357
Address: http://www.ncbi.nlm.nih.gov/pubmed/22355202
Date Issue: 2011
Appears in Collections:Artigos e Materiais de Revistas Científicas - Unicamp

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